Preliminary report of in vitro and in vivo effectiveness of dornase alfa on SARS-CoV-2 infection.

Dornase alfa, the recombinant form of the human DNase I enzyme, breaks down neutrophil extracellular traps (NET) that include a vast amount of DNA fragments, histones, microbicidal proteins and oxidant enzymes released from necrotic neutrophils in the highly viscous mucus of cystic fibrosis patients. Dornase alfa has been used for decades in patients with cystic fibrosis to reduce the viscoelasticity of respiratory tract secretions, to decrease the severity of respiratory tract infections, and to improve lung function. Previous studies have linked abnormal NET formations to lung diseases, especially to acute respiratory distress syndrome (ARDS). It is well known that novel coronavirus disease 2019 (COVID-19) pneumonia progresses to ARDS and even multiple organ failure. High blood neutrophil levels are an early indicator of COVID-19 and predict severe respiratory diseases. Also it is reported that mucus structure in COVID-19 is very similar to that in cystic fibrosis due to the accumulation of excessive NET in the lungs. In this study, we showed the recovery of three individuals with COVID-19 after including dornase alfa in their treatment. We followed clinical improvement in the radiological analysis (two of three cases), oxygen saturation (Spo2), respiratory rate, disappearance of dyspnoea, coughing and a decrease in NET formation and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral load after the treatment. Also here, we share our preliminary results suggesting that dornase alfa has an anti-viral effect against SARS-CoV-2 infection in a green monkey kidney cell line, Vero, and a bovine kidney cell line, MDBK, without determined cytotoxicity on healthy peripheral blood mononuclear cells.

Effects of imatinib mesylate on renin-angiotensin system (RAS) activity during the clinical course of chronic myeloid leukaemia.

The renin-angiotensin system (RAS) is involved in cell growth, proliferation and differentiation in bone marrow in an autocrine-paracrine manner, and it modulates normal and neoplastic haematopoietic cell proliferation. This study aimed to assess expressions of the RAS components, renin, angiotensinogen and angiotensin-converting enzyme (ACE), during imatinib mesylate treatment of patients with chronic myeloid leukaemia (CML). Expressions of RAS components were studied in patients with CML at the time of diagnosis (n = 83) and at 3, 6 and 12 months after diagnosis (n = 35) by quantitative real-time polymerase chain reaction. De novo CML patients had increased ACE, angiotensinogen and renin mRNA levels and these expression levels decreased following administration of imatinib. The RAS activities were significantly different among Sokal risk groups of CML, highlighting the altered biological activity of RAS in neoplastic disorders. The results of this study confirm that haematopoietic RAS affects neoplastic cell production, which may be altered via administration of tyrosine kinase inhibitors such as imatinib mesylate.

The effects of heparin on DLD-1 colon cancer cell line.

BACKGROUND: Patients with cancer are frequently treated with anticoagulants, including heparin, to prevent or to treat the deep vein thrombosis. It has been indicated that heparin affects the survival of patients with cancer. Also, the apoptotic and antiproliferative effects of heparin on some cancers has been demonstrated. Experimental studies support the hypothesis that cancer progression can be influenced by heparin, but the results of these studies are not conclusive. OBJECTIVES: We planned to investigate the effects of different concentrations of heparin in the colon cancer cell line DLD-1. METHODS: This study was done by the addition of heparin in different doses into colon cancer cell line DLD-1 in vitro. After an incubation period of 72 hours, study and control groups were evaluated for viable cell count, percentage of proliferating index and apopitosis percentage. RESULTS: The result of the viable cell count in the second and third study groups (98.35+/-27.3, 97.23+/-39.38) were low compared to the control group. The results of the proliferative index of study groups (46.47+/-10.44, 47.23+/-12.03, 45.55+/-14.2) were higher than the control group (40.62+/-9.28). The results of apoptosis in study groups (14.35+/-1.93, 16.47+/-7.25, 13.56+/-5.66) were lower compared to the control group (22.17+/-15.9). But these results were not statistically significant. CONCLUSION: Heparin has no significant anti-proliferative and apoptotic effects on colon cancer cells in vitro. Therefore, for clinical applications, more advanced studies are needed to examine the effect of heparin on colon cancer (Tab. 3, Fig. 3, Ref. 25). Full Text (Free, PDF) www.bmj.sk.

A detachment technique based on the thermophysiologic responses of cultured mesenchymal cells exposed to cold.

Trypsinization has generally been used as a technique to detach adherent mesenchymal stromal cells (MSC). However, this technique involves chemical manipulation. This study was designed to identify whether detachment of MSC can be induced by cold without using trypsin. MSC isolated from bone marrow were detached via trypsin or exposed to -20 degrees C for 1, 5 or 10 min at all passages. Compared with trypsinization, exposing MSC to -20 degrees C for 10 min resulted in a significant decrease in MSC number and viability. In conclusion, although detachment of adhered MSC on culture dishes via exposure to cold may allow structurally and functionally intact detached cells, the technique requires improvement of the thermotolerance of MSC.

Active immunotherapy for cancer patients using tumor lysate pulsed dendritic cell vaccine: a safety study.

Cancer vaccine therapy represents a promising therapeutical option. Consistently, with these new treatment strategies, the use of dendritic cell vaccines is becoming increasingly widespread and currently in the forefront for cancer treatment. The purpose of this study was to evaluate the feasibility and safety of tumor lysate-pulsed dendritic cell (DC) vaccine in patients with advanced cancers. For this purpose, eighteen patients with relapsed or refractory cancer were vaccinated with peripheral monocyte-derived DCs generated with GM-CSF and IL-4, and pulsed consequently with 100 microg/ml of tumor lysate before maturation in culture in the presence of IL-1beta, PGE2 and TNF alpha for two days. The first two vaccinations were given intradermally every two weeks while further injections were given monthly. Tumor lysate-pulsed dendritic cell injections were well-tolerated in all patients with no more than grade 1 injection-related toxicity. Local inflammatory response was mainly erythematous which subsided in 48 hrs time. No end organ toxicity or autoimmune toxicity was identified. Clinical responses observed in our study were satisfactory for a phase I clinical study. We observed 4 (22%) objective clinical responses. These responses are significantly correlated with delayed type hypersensitivity testing (DTH) (p < 0.01). The results showed that this active immunotherapy is feasible, safe, and may be capable of eliciting immune responses against cancer.

HLA alleles and haplotypes in the east Black Sea Turkish population.

HLA class I and class II alleles were studied for the first time in 234 unrelated individuals inhabiting the East Black Sea region in Turkey. This region is on the historic silk road and close to Georgia. HLA class I (A* and B*) and class II DRB1* typing was done by the PCR-SSP method. A total of 17 HLA-A* alleles, 34 HLA-B* alleles, and 15 HLA-DRB1* alleles were detected. HLA-A*-B*, A*-DRB1*, and B*-DRB1* two-loci linkage disequilibrium data show that two specific combinations (A2-B35, A2-DRB1*11, and B35-DRB1*13) had the highest frequency (more than three or four times) compared with the other two-loci combinations, possibly reflecting an ancient founder effect. A*24 B*18 DRB1*13 and A*32 B*27 DRB1*11 were the most common haplotypes in the east Black sea Turkish population. HLA-B* showed the highest heterozygosity (94%) among the samples. The observed diversity in the HLA-A* and HLA-DRB1* loci was quite similar, ranging from 79% to 84%. We suggest that the east Black Sea Turkish population is characterized by the features of the Turkish anthropological type with some influence of other groups, such as Caucasians, Asians, and Mediterraneans.

The in-vitro effects of medroxyprogesterone acetate on acidic pH induced apoptosis of periferal blood mononuclear cells.

Medroxyprogesterone acetate (MPA) which has a steroid structure, is widely used in oncology practice in the treatment of the cachexia of cancer and to reduce hematologic toxicity in patients receiving chemotherapy. However, the mechanisms of MPA on these effects are unclear. In this study, we investigated the effects of two different doses of MPA (10(-5) and 10(-6) M/L) on acidic pH induced apoptosis of periferal blood mononuclear cells (PBMC) derived from 10 healthy volunteers. Compared with the control group, we found that MPA at the dose of 10(-5) M/L had a negative effect on apoptosis (32.88 +/- 4.61 and 20.7 +/- 1.53% respectively, p < 0.05), a positive effect on cell count of PBMC (1395 +/- 151 x 10(3) and 1100 +/- 139 x 10(3) respectively, p < 0.05) and no effects on cell viability and its proliferation. More comprehensive trials are needed to clarify this effect of MPA.

Philadelphia negative, Bcr-Abl positive chronic myeloid leukemia associated with pure red cell aplasia.

Pure red cell aplasia (PRCA) is a rare disorder which is associated with thymoma, viral infections and autoimmune diseases. A few cases of PRCA during the clinical course of CML have been reported and these usually terminate in blastic crisis and death, suggesting a poor prognosis. However, only one case of Philedelphia chromosome negative, Bcr-Abl positive CML associated with PRCA has been reported. Here, we present a second case report of a Philedelphia negative, Bcr-Abl positive CML associated with PRCA who was unresponsive to all the chemotherapeutic regimens. We conclude that the present case supports the idea that the development of PRCA in the course of CML may be a bad prognostic sign.

The effects of dexamethasone on leukemic cells derived from patients with AML.

This study was performed to investigate the effects of dexamethasone on acute myelocytic leukemia (AML) cells. The study has been designed to investigate the in vitro effects of dexamethasone on the proliferative capacity, maturation and apoptosis of leukemic cells derived from patients with AML. Dexamethasone induced leukemic cell apoptosis and decreased cell count compared to the control. The percentage of cellular maturation increased in the dexamethasone groups compared to the control groups. Proliferative capacity of the cells was similar in all groups. These effects of dexamethasone on leukemic cells may be related to activation of the genes which stimulate apoptosis and maturation. More comprehensive studies are needed to evaluate the apoptotic effect of dexamethasone on AML cells.

Effect of interferon-alpha(2a) on neutrophil adhesion and phagocytosis in chronic myeloid leukemia and Behçet's disease.

Alpha-interferon (alpha-IFN) is implicated in a Behçet's disease (BD)-like syndrome observed in a small number of chronic myeloid leukemia (CML) patients. The effect of alpha-IFN on neutrophil adhesion and phagocytosis in CML patients, BD patients and healthy volunteers was investigated to clarify the reason for this observation. Ten subjects were studied for each group by incubating neutrophils with various doses of alpha-IFN. Basal neutrophil adhesions for CML patients, BD patients and healthy volunteers were similar. However, BD patients had greater basal phagocytosis than CML patients, and both groups had greater basal phagocytosis than healthy volunteers. Neutrophil adhesion and phagocytosis of CML patients increased following incubation with higher doses of alpha-IFN, and phagocytosis approached the high levels observed with BD neutrophils. This study provides evidence that alpha-IFN activates neutrophils in CML patients in a dose-dependent manner, and leads to a neutrophil function profile that resembles BD.

Factor V Leiden and its relation to left ventricular thrombus in acute myocardial infarction.

OBJECTIVE: The genetic defect of coagulation factor V, known as factor V Leiden, produces a resistance to degradation by activated protein C (APC) and increases the risk of venous thrombosis. However, the role of factor V Leiden in the formation of left ventricular (LV) thrombus has not been studied. We investigated whether factor V Leiden is a risk factor for LV thrombus in patients with acute myocardial infarction (AMI). METHODS AND RESULTS: We have analyzed clinical, echocardiographic and biochemical data in 135 consecutive patients (aged 58 +/- 13 years; 31 women) with first anterior AMI. Two-dimensional echocardiographic examination was performed on days 1, 3, 7, 15 and 30; LV thrombus was detected in 33 (24.4%) of 135 patients with AMI. The study also included 95 control subjects. Healthy age and sex-matched subjects without a personal or family history of ischaemic heart disease, stroke or thromboembolic disease served as a control group. Blood samples from the patients and controls were analyzed for the factor V Leiden mutation by DNA analysis, using the polymerase chain reaction. In addition, concentrations of fibrinogen, von Willebrand factor (vWF), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1) and D-dimer were measured in 135 patients. There was no significant difference in the prevalence of factor V Leiden between patients and control subjects. The prevalence of the factor V mutation was 9% (3/33) in patients with thrombus, and 7.7% (8/103) in patients without thrombus. The prevalence of factor V Leiden was 7.3% (7/95) in control subjects. No significant differences in plasma fibrinogen (480 +/- 195 vs. 444 +/- 179 mg/dl, p = 0.6), D-dimer (471 +/- 256 vs. 497 +/- 293 ng/dl, p = 0.7), vWF (112 +/- 18 vs. 103 +/- 15%, p=0.5), PAI-1 (26.7+/- 9.8 vs. 28.1 +/- 10.2 ng/dl, p = 0.6), and t-PA (19.8 +/- 8.7 vs. 17.2 +/- 9.1 ng/dl, p = 0.7), levels are found in patients with LV thrombus when compared with those without LV thrombus. Multivariate analyses showed that peak creatine kinase level (p = 0.002) and LV wall motion score index (p = 0.003) were independent predictors of LV thrombus formation. CONCLUSION: Factor V Leiden mutation is not a risk factor for LV thrombus formation in patients with AMI.

[Effects of factor V Leiden mutations on prognosis in patients with acute myocardial infarction]. / Akut miyokard infarktüslü hastalarda faktör V Leiden mutasyonunun prognoz üzerine etkisi.

OBJECTIVE: Factor V Leiden mutation, the genetic defect underlying resistance to activated protein C, is the most common risk factor for venous thrombosis. Factor V Leiden mutation and its relation to post-myocardial infarction (MI) complications including angina pectoris, heart failure, reinfarction and cardiac mortality has not been investigated. We aimed to investigate this relation. METHODS: The prevalence of factor V Leiden mutation was investigated in 122 patients with first acute myocardial infarction (aged 56 +/- 11, 82 men/40 women). These patients were divided into two groups according to whether the patients had factor V Leiden mutation (Group I) or not (Group II). Post MI complications were evaluated during 18 months. Blood samples from the patients were analyzed for factor V Leiden mutation by DNA analysis, using the polymerase chain reaction (PCR). RESULTS: Factor V Leiden was detected in 11 (9%) patients (aged; 54 +/- 10, 5 women/men) and was not detected in 111(90%) patients (aged; 56 +/- 11; 35 women/76 men) of the 122 patients. There were no significant differences between Group I and Group II in terms of post MI complications, including reinfarction (27% vs. 29%; p > 0.05, respectively), angina pectoris (45% vs. 38%; p > 0.05, respectively), heart failure (27% vs. 23%; p > 0.05, respectively) and cardiac mortality (18% vs. 14%; p > 0.05, respectively). CONCLUSION: Post MI complications, including reinfarction, heart failure, angina pectoris and cardiac mortality were not increased in patients with factor V Leiden.

National survey of hemapheresis practice in Turkey (1998).

The Turkish Apheresis Group has maintained a national registry for apheresis activities since 1997. The hemapheresis practice of Turkey in 1998 is summarized in brief detail in this article. A total of 30, 136 apheresis procedures were performed at 31 different apheresis centers. At 10 centers, 145 peripheral blood stem cell (PBSC) apheresis were performed on 82 patients in allogeneic setting and at 17 centers, 981 PBSC apheresis were performed on 271 patients in autologous setting. Frequently observed adverse effects during PBSC apheresis were mild tremor and chills, paresthesia and nausea in 15% of the patients and donors. Vascular access complications, particularly observed in autologous setting due to central venous catheters were encountered in 10% of the procedures. Eight hundred and sixty-nine therapeutic plasma exchange procedures were performed at 21 centers on 172 patients, most commonly for neurological disorders and thrombotic thrombocytopenic purpura (TTP)/hemolytic uremic syndrome (HUS). Therapeutic cytapheresis procedures like leukapheresis, plateletapheresis and erythrocyte apheresis were performed especially for cytoreduction in myeloproliferative disorders. A total of 204 cytapheresis procedures (66% leukapheresis, 33% plateletapheresis and 1% erythrocytapheresis) were performed on 134 patients in 15 centers. Donor plateletapheresis was the most used apheresis procedure, reaching a total of 28.016 in 1998. Many university hospitals and a few state hospitals are performing above-mentioned apheresis procedures with great success and acceptable side effects. According to these data we are planning prospective trials and will establish National Standards of Practice.

Role of hepatocyte growth factor in the development of dendritic cells from CD34+ bone marrow cells.

BACKGROUND AND OBJECTIVE: Hepatocyte growth factor (HGF) is known to augment the effects of stem cell factor, interleukin-3, granulocyte-macrophage colony-stimulating factor (GM-CSF), erythropoetin, and granulocyte colony-stimulating factor, all of which are involved in hematopoiesis. HGF is also known to have a role in immune responses. The aim of this study was to investigate whether HGF is involved in the development of dendritic cells (DC) from CD34+ bone marrow cells. DESIGN AND METHODS: CD34+ cells obtained from three healthy donors were incubated in various combinations of HGF, GM-CSF, and tumor necrosis factor (TNF) for 12 days. Developing cell populations were analyzed for surface markers, morphology and functional capacities by flow cytometry, light microscopy and mixed lymphocyte reaction, respectively. RESULTS: Incubation with HGF alone generated greater number of dendritic cells from CD34+ bone marrow cells than incubation with GM-CSF, or a combination of GM-CSF with TNF. HGF was also found to potentiate the effect of GM-CSF on DC and monocyte development. The effects of HGF were inhibited by the concurrent use of TNF. INTERPRETATION AND CONCLUSIONS: HGF appears to be a significant factor in the development of dendritic cells from CD34+ bone marrow cells.

The in vitro effects of isoflurane, sevoflurane, and propofol on platelet aggregation.

UNLABELLED: We studied the in vitro effects of sevoflurane, isoflurane, and propofol anesthesia on platelet function. Thirty patients undergoing minor surgical procedures were divided into three groups (n = 10 each). Induction of anesthesia was achieved by using 5 mg/kg thiopental i.v., and 0.1 mg/kg vecuronium i.v. was used for muscle relaxation. Anesthesia maintenance was provided by sevoflurane in the first, isoflurane in the second, and propofol infusion in the third group with 70% N2O in O2. Hemoglobin, hematocrit, thrombocyte count, prothrombin time, activated partial thromboplastin time, international normalized ratio, arterial pH, von Willebrand factor, viscosity, platelet aggregation, and bleeding time were measured 1 h pre-, intra-, and postanesthesia. There was no difference among the platelet aggregation ratios of the pre-, intra-, and postoperative periods in the isoflurane group. The aggregation ratios in the sevoflurane and propofol groups were significantly reduced at intraoperative periods compared with preoperative values. Diminished aggregation values were also found 1 h postoperatively compared with the control values in the sevoflurane and propofol groups. We conclude that, in patients with a bleeding tendency during the intra- and early postoperative period, isoflurane may be preferred as a general anesthetic. IMPLICATIONS: In our study, using vacuum-operated tubes, we demonstrated that sevoflurane and propofol had a significant inhibitory effect on intraoperative and early postoperative platelet aggregation, whereas isoflurane had no effect. Therefore, isoflurane may be preferred as a general anesthetic in patients with a clinically relevant bleeding tendency.

The inhibiting effect of cola on gastric mucosal cell cycle proliferation in humans.

BACKGROUND: Acidic beverages may be involved in regulating the cell proliferation of the gastric mucosa. We therefore analyzed the interaction of Coca-Cola consumption and gastric mucosal proliferation by means of flow cytometry. METHODS: Sixteen healthy students agreed to participate in this study. All volunteers underwent an oesophagogastroduodenoscopy after a 12-h overnight fast. Endoscopic changes in the gastric mucosa were determined quantitatively. One day later, after a 12-h overnight fast, all volunteers received standard Coca-Cola (200 ml, pH 2.6, 4 degrees C). One hour later all volunteers again underwent oesophagogastroduodenoscopy, to measure gastric mucosal damage. During both the first and the second endoscopy at least four biopsy specimens were taken from the antrum for flow cytometric analysis. RESULTS: The endoscopic analysis showed that there was no difference before and after Coca-Cola consumption. However, the flow cytometric analysis showed that Coca-Cola inhibited the proliferation index and the S phase. Before Coca-Cola consumption G0/G1: 60 (57-62), G2/M: 0.6 (0.2-1), S: 40 (37-42), and PI: 0.40 (0.38-0.43) and after Coca-Cola consumption G0/G1: 70 (60-73), G2/M: 1.9 (1.2-2.5), S: 28 (26-32), and PI: 0.30 (0.27-0.34) the cell population G0/G1 and G2/M phases were significantly increased (P < 0.0001, 0.0003), and the cell population S and PI phases were significantly low compared with the pre-consumption data (P < 0.0002, 0.0001). CONCLUSION: The cell cycle analysis reflects that Coca-Cola inhibits a crucial event in the cell cycle occurring at the G1/S border.

Effect of long-acting somatostatin analogue (octreotide, SMS 201-995) plus calcium channel blocker (verapamil) on gallbladder contraction.

BACKGROUND/AIMS: In this study we investigated the effect of the long-acting somatostatin analog octreotide (SMS 201-995) plus calcium channel blocker (Verapamil) on gallbladder contraction. METHODOLOGY: Fourty healthy volunteers participated in this study. Gallbladder volumes were measured by ultrasonography. After recording the baseline measurement, the volunteers received either saline (n:10), or SMS 201-995 100 B microgram subcutaneously (s.c.) (n:10) or verapamil 80 mg peroral (po) (n:10), or verapamil plus SMS 201-995 (n:10). Two hours later the gallbladder volumes were rescanned in 15 min intervals for 60 min. At the end all volunteers received standard liquid test meal (ensure 250 Cal/250 ml) and scans were again performed for one hour. RESULTS: The mean baseline gallbladder volume was 18.6 +/- 5.2 ml in all groups. The gallbladder volumes in the placebo group were 18.6 +/- 5.2 to 19.0 +/- 10.2 ml. In this group, after administration of test meal decreased the mean gallbladder volume to 14.3 +/- 7.5 to 8.4 +/- 5.8 ml, but these values were not significantly different from the baseline values. In the verapamil group the volumes increased from 18.6 +/- 5.2 to 28.5 +/- 9.7 to 30.8 +/- 11.6 ml. These values were significantly different from the baseline and the control group (p < 0.05). In this group, post-prandial mean volumes decreased to baseline in 30 min, but these values were higher than in the placebo group (p < 0.01). Verapamil-induced fasting the gallbladder relaxation was totally abolished to the placebo value by SMS 201-995. In verapamil plus SMS 201-995 and SMS 201-995 alone groups, the fasting and post-prandial volumes did not change when compared to the baseline value, but post-prandial volumes were higher than the placebo (p < 0.01). CONCLUSION: These results suggest that verapamil-induced gallbladder relaxation was totally abolished by SMS 201-995.

Effect of high-dose medroxyprogesterone acetate on tumor necrosis factor-alpha release in patients with chemotherapy-induced neutropenia.

The aim of the study was to investigate the effects of high-dose medroxyprogesterone acetate (MPA) on the tumor necrosis factor-alpha (TNF-alpha) release in patients with chemotherapy-induced neutropenia. We also evaluated the effects of high-dose MPA on hematological parameters (leukocyte, neutrophil, platelet, hemoglobin, hematocrit) and side effects of MPA. One week following the first cycle chemotherapy, 20 patients who developed neutropenia were enrolled in the study. One gram/day MPA was administered orally to the patients and was continued from one week following the first chemotherapy cycle to one week after the second chemotherapy cycle. The patients received the second chemotherapy cycle at the same dosages as the first cycle. Before MPA treatment TNF-alpha levels were lower than post-treatment levels, but the difference was not statistically significant (P > 0.05). The differences in the mean leukocyte and neutrophil counts before and after the high-dose MPA treatment were statistically significant (p < 0.05).